Fig 2: Impact of maternal Tdap-IPV vaccination on maternal innate cytokine responses to B. pertussis. Maternal blood was treated for 24 h with heat-killed B. pertussis in duplicate and cytokines were quantified in supernatants in singlicate. Data points represent cytokine concentrations plotted on a log10 scale. Unvaccinated n=10 (filled blue circles); vaccinated n=11 (open green circles). Vaccinated and unvaccinated groups were compared by Student's t test (normal distribution; IL-6 and IL-10) or Mann-Whitney U test (non-normal distribution; all others); * p<0.05, ** p<0.01. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Fig 3: Changes in cytokine production by lymphocytes during acute and convalescent COVID-19(A–C) Graphs showing frequencies of CD4+ T cells that stain positive for (A) IL-10, (B) IL-17, and (C) IFNγ and TNF-α following 3-h stimulation with PMA and ionomycin in healthy individuals (n = 25–30), acute COVID-19 patients (n = 29–33), and convalescent COVID-19 patients with normal (n = 55–57) or abnormal chest X-ray findings (n = 25–26).(D) Graphs showing frequencies of CD8+ T cells that stain positive for IFNγ and TNF-α following 3-h stimulation with PMA and ionomycin in healthy individuals (n = 28), acute COVID-19 patients (n = 24–31), and convalescent COVID-19 patients with normal (n = 54–57) or abnormal chest X-ray findings (n = 21–24).(E and F) Graphs show frequencies of (E) CD4+ and (F) CD8+ T cells that stain positive for IFNγ and TNF-α following 3-h stimulation with PMA and ionomycin in convalescent COVID-19 patients who initially presented with mild (n = 13–14), moderate (n = 25–28), and severe (n = 34–41) disease.(G and H) Graphs track frequencies of (G) CD4+ and (H) CD8+ T cells that stain positive for IFNγ and TNF-α in the same COVID-19 patient at acute (gray circles) and convalescent (maroon circles) time points (n = 14).(I) Graphs showing frequencies of CD19+ B cells positive for IL-10, IL-6, and TNF-α following 48-h stimulation with CpGB in healthy individuals (n = 22–27), acute COVID-19 patients (n = 22–32), and convalescent COVID-19 patients with normal (n = 52–54) or abnormal chest X-ray findings (n = 24–27).(J) Graphs track frequencies of CD19+ B cells that stain positive for IL-10, IL-6, and TNF-α in the same COVID-19 patient at acute (gray circles) and convalescent (maroon circles) time points (n = 11–14). Graphs show individual patient data, with the bar representing median values.In all graphs, open triangles represent SARS-CoV-2 PCR− patients. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, 1-way ANOVA with Kruskal-Wallis test with Dunn’s post hoc testing for multiple comparisons, except for graphs showing CD4+TNF-α+ and CD8+IFNγ+ T cells in (E) and (F), where 1-way ANOVA with Holm-Sidak post hoc test was used, or Wilcoxon matched-pairs signed rank test (G, H, and J).See also Figures S3 and S4.

Fig 4: Metformin inhibited the production of proinflammatory cytokine IL-6 in CD3 negative cells. Representative flow analysis of IL-6 production and MFI level in CD3- cells (a), CD3-CD57 + cells (c) and CD3-CD57- cells (d) between 20 mM Met treatment and control for 24 h in middle age group. Representative flow analysis of IL-6 production and MFI in CD3-CD57 + and CD3-CD57- population (b). Expressed as the mean ± SEM. ***P < 0.001; Paired t test. Met, metformin

Fig 5: Variations in double-positive (IL-6 and TNF-α positive) monocytes and mDCs after dia. E. coli and TLR stimulation on days 14 and 35.A significant increase in the percentage of monocytes that were simultaneously producing IL-6 and TNF-α was seen after ex vivo stimulation with LPS (200 ng/mL), flagellin (500 ng/mL), or dia. E. coli (1E7 CFU/well) on day 35, comparing to day 14 (A). The average percentage of double-positive monocytes was almost doubled after stimulation with all stimuli in contrast to the RPMI control (B). Following the stimulation with the same stimuli, mDCs showed a significant increase in IL-6 and TNF-α production after exposure to LPS, flagellin, and dia. E. coli (C). Just about all participants showed an increase in double-positive mDCs after dia. E. coli and LPS stimulation, with an average percentage increase of almost 1.5 times (D). For statistical analysis, paired sample t-test was applied. Data are represented in Whisker-plots with a median, 25% and 75% quartile. * p<0.05; ** p<0.01; *** p<0.001,**** p<0.0001. A visual representation of changes in the percentage of IL-6 and TNF-α producing monocytes (E) and mDCs (F) of a representative participant (#10) on day 14 compared to day 35. The percentage of double-positive (top-right quadrant) monocytes increased from 14.3% to 49% (E), whereas double-positive mDCs had an increase from 18.1% on day 14 to 33% on day 35 (F) in this selected participant.